In vitro and in vivo interactions between SMG7 and p53. ( a ) Scheme of human SMG7 protein containing the N-terminal 14-3-3-like Tetratricopeptide repeat (TPR) and C-terminal low complex region. ( b ) Immunofluorescence microscopy of H1299 cells transiently expressing FH-SMG7 stained with α-HA antibody (green) and 4,6-diamidino-2-phenylindole (DAPI; blue). ( c ) Cell extracts from H1299 cells transiently expressing p53 with or without FH-SMG7 and the α-Flag immunoprecipitates were analyzed using western blot using α-SMG7 and α-p53 antibodies. ( d , e ) SMG7 binding to p53 in vitro . GST or GST-p53 fusion proteins (full length or various fragments) were used in pulldown assays with purified FH-SMG7 ( d ) or in vitro translated FH-SMG7 ( e ). FH-SMG7 was detected by western blot and GST proteins stained with Ponceau S. ( f , g ) Endogenous interaction of SMG7 with p53 in HCT116 ( f ) and U2OS cells ( g ). Cell extracts and the α-normal mouse IgG or α-p53 immunoprecipitates were analyzed with western blot analysis using α-SMG7 and α-p53 antibodies. ( h ) Cell extracts from control or irradiated cells (at various time points) and the α-Flag immunoprecipitates were analyzed with western blot analysis using antibodies as indicated.
{"Image ImageWidth": "1868", "Image ImageLength": "4122", "Image BitsPerSample": "[8, 8, 8]", "Image Compression": "LZW", "Image PhotometricInterpretation": "2", "Image Orientation": "Horizontal (normal)", "Image SamplesPerPixel": "3", "Image XResolution": "300", "Image YResolution": "300", "Image PlanarConfiguration": "1", "Image ResolutionUnit": "Pixels/Inch", "Image Software": "Adobe Photoshop CS4 Windows", "Image DateTime": "2015:11:13 12:36:16", "Image ExifOffset": "252", "Thumbnail Compression": "JPEG (old-style)", "Thumbnail XResolution": "300", "Thumbnail YResolution": "300", "Thumbnail ResolutionUnit": "Pixels/Inch", "Thumbnail JPEGInterchangeFormat": "390", "Thumbnail JPEGInterchangeFormatLength": "5253", "EXIF ColorSpace": "Uncalibrated", "EXIF ExifImageWidth": "1050", "EXIF ExifImageLength": "2317"}